Immunological Detection of Fusarium Species in Cornmeal

Abstract

An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect Fusarium species in foods. Antibodies to proteins extracted from the mycelia of Fusarium graminearum and Fusarium moniliforme (verticillioides) were produced in New Zealand white rabbits. These antibodies detected 13 Fusarium species in addition to the producer strains. Levels of Fusarium semitectum and Fusarium tricinctum strains were below the detection threshold. The specificity of the assay was tested against 70 molds and yeasts belonging to 23 genera. One strain of Monascus species and one strain of Phoma exigua were detected; however, these two molds are not common contaminants of cereal grains or foods and should not interfere with the assay. The indirect ELISA's detection limits for F. graminearum and F. moniliforme were 0.1 and 1 microg of mold mycelium per ml of a cornmeal mixture, respectively. When spores of each mold were added individually to cornmeal mixtures (at ca. 10 spores per g) and incubated at 25 degrees C, these spores were detected by the indirect ELISA when they reached levels of 10(2) to 10(3) CFU/ml after 24 to 36 h. The indirect ELISA developed here shows promise for the detection of Fusarium species in grains or foods.

Evaluation of arbuscular mycorrhiza and other biocontrol agents in managingFusarium oxysporumf. sp.Cubenseinfection in banana cv. Neypoovan

“…The polyclonal antibody developed by us was able to detect the Fusarium in plants before the appearance of the visual symptoms though careful standardization of the technique for large scale use was not carried out. Iyer and Cousin (2003) used similar techniques to study Fusarium infection of corn grains. In our studies the antibodies did not cross react with AM or Trichoderma species.…”

Development of polyclonal antibodies againstFusarium verticillioidesexoantigens

“…Tsai and Yu (1997) reported a double-sandwich ELISA for A. parasiticus and A. flavus with detection limit of 1 mg/ml. Iyer and Cousin (2003) developed an indirect ELISA for F. graminearum and F. verticillioides with detection limits of 0.1 and 1 mg mould mycelium/ml corn meal mixture, respectively.…”

“…The antibodies to F. poae were species specific although those for F. sporotrichioides and F. graminearum were genera specific, showing cross-reactivity of 63% and 55%, respectively with F. verticillioides exoantigens. Iyer and Cousin (2003) raised antibodies in rabbits against mycelia proteins from F. verticillioides and F. graminearum and analysed the cross-reactivity with moulds and yeasts belonging to 23 genera. Among the moulds tested, Monascus sp., Phoma exigua , and most Fusarium species showed crossreactivities higher than 30%, considered positive for Fusarium antibody.…”

Detection ofFusariumsp. contamination in corn by enzyme-linked immunosorbent assay

“…The development of the enzyme-linked immunosorbent assay (ELISA) (Engvall & Pearlmann 1972) could be a promising alternative to the traditional methods (Lin et al 1986). Several ELISAs have been developed for detecting and identifying moulds with variable success (Notermans et al 1986, Tsai & Yu 1997, Chen et al 2000, Yong & Cousin 2001, Eibel et al 2005, Yucel et al 2005, however a limited number have been developed for the detection of Fusarium species in foods (Gan et al 1997, Abramson et al 1998, Iyer & Cousin 2003, Park et al 2003.…”